酸奶弱后酸化发酵剂乳酸菌育种技术研究进展

后酸化是影响酸奶贮藏期间感官品质和风味特性的重要因素,是决定产品货架期的重要指标。弱后酸化发酵剂菌种的选育是优良发酵剂开发的重要技术手段。通过诱变育种筛选出具有弱后酸化特性的菌株,是延缓酸奶贮藏期间后酸化的技术途径之一。国内外围绕弱后酸化发酵菌株选育,采用诱变育种方法开展了大量卓有成效的研究。本研究针对诱变育种的方法、原理与研究现状,结合本课题组近几年关于发酵剂菌种弱后酸化的研究工作,对弱后酸化乳酸菌菌种的诱变育种进行了简要阐述与展望。     

泛谈酸奶

酸奶(Yoghurt)又名酸乳或酸凝乳。它是利用消毒处理后的鲜奶为培养基加入发酵剂——乳酸菌制成的。酵解使鲜乳中的乳糖变为乳酸,鲜乳中的蛋白质遇酸而凝固,形成酸化酪蛋白,即酸奶。人类制做酸奶已有几千年的历史,古老的方法是自然发酵。在二十世纪初,俄国著名生物学家米奇尼柯夫发现,保加利亚有个“长寿村”,百岁老人很多,重要     

酸奶发酵剂和乳酸菌生物技术育种

由新鲜牛奶制成的酸奶由于其丰富的营养、特殊的风味、爽滑的质构和良好的生理功能 ,倍受人们青睐。对酸奶乳酸菌的微生物种类、功能及其组成的酸奶发酵剂进行了综述 ,并简要介绍了乳酸菌生物工程育种方面的研究进展及酸奶乳酸菌研究领域一些令人感兴趣的课题。     

保存温度对酸奶品质及活性乳酸菌含量的影响

以2种市售酸奶作为研究对象,研究不同保存温度(4、28、37℃)对酸奶品质及活性乳酸菌含量的影响,并分析酸奶酸度、pH值及活性乳酸菌含量的变化趋势。实验结果表明,保存温度为4℃时,酸奶的pH值和酸度具有较高的稳定性及较高的活性乳酸菌含量,但其稳定性却比保存温度为28和37℃时差,很容易发生乳清分离现象。     

斑马鱼功效模型在验证乳制品补钙和促进肠蠕动中的应用

目的基于斑马鱼补钙和肠蠕动模型,验证和对比不同牛奶补钙和酸奶促进肠蠕动的作用效果。方法选择发育健康且阶段一致的3 dpf野生型斑马鱼,加入不同浓度(2. 5、5. 0、10. 0 mg/m L)的纯牛奶稀释液,同时设置空白对照组。处理至7 dpf后,用2 mg/m L钙黄绿素染色,在荧光显微镜下观察拍照,计算斑马鱼的椎骨荧光强度。选取5 dpf的斑马鱼,用尼罗红染料处理16 h,加入不同浓度(2. 5、5. 0、10. 0 mg/m L)的酸奶和乳酸菌饮料稀释液,同时设置空白对照组。处理24 h后,在荧光显微镜下观察拍照,计算斑马鱼肠道内荧光强度。结果纯牛奶、酸奶和乳酸菌饮料作用效果都随浓度的增加而增强。与空白对照组比较,6种纯牛奶中2种纯牛奶处理的斑马鱼骨骼荧光强度有一定增加; 8种酸奶和乳酸菌饮料都显著引起了肠道荧光强度的减弱,酸奶的效果优于乳酸菌饮料。结论纯牛奶、酸奶和乳酸菌饮料对斑马鱼的最大耐受浓度(MTC)为10 mg/m L,且对斑马鱼起到补钙和促进肠蠕动的作用。     

酸奶中乳酸菌含量的检测

目的检测酸奶发酵前后以及贮藏期间的乳酸菌活菌数及酸度变化,以对酸奶的质量进行控制.方法应用高层半固体琼脂试管法. 结果酸奶中乳酸菌含量极高,达2.65×1010个/ml以上,并随贮藏时间的延长而发生变化. 糖对酸奶的含菌量无明显影响.双菌酸奶比单菌酸奶含菌量高,常可达到倍增,pH值随发酵和贮藏进程而下降.结论高层半固体琼脂试管法操作简便、快速、准确、活菌检出率高.     

弱后酸化瑞士乳杆菌的筛选及发酵特性研究

[目的]在大健康背景下,发酵乳凭借其优良的营养和保健功能备受青睐.后酸化是发酵乳在贮存、运输和销售等过程中活性乳酸菌继续代谢产酸导致的,严重影响产品的感官品质,降低菌株的存活率.本文通过筛选弱后酸化的瑞士乳杆菌,改善发酵乳的后酸化问题.[方法]本研究以瑞士乳杆菌L551为出发菌株,硫酸新霉素为筛选压力,筛选弱后酸化的瑞...     

酸奶能够克服“乳糖不耐受”的原因探究

随着人们生活水平的提高和对营养健康的关注,牛奶成了家庭必备的食品。但有些人喝牛奶会腹胀、腹泻,而喝酸奶会避免这个症状。通过上网查阅资料,采访专家,参考文献,开展社会调研,亲自制作酸奶,实地考察酸奶生产线,以及对对酸奶成分进行检测后,发现喝牛奶腹胀、腹泻的现象(乳糖不耐受)较为普遍,其主要原因是体内缺乏乳糖酶的人群不能分解牛奶中的乳糖所致。酸奶与牛奶的加工工艺不同,在发酵过程中20%-30%的乳糖被分解,含量明显下降,易于人体消化吸收。加之酸奶中的活性乳酸菌的代谢物可增加小肠乳糖水平,从而有效缓解和克服了乳糖不耐受的症状。     

绿豆酸奶的制作工艺

绿豆酸奶是以绿豆为主要原料制作的一种新型植物蛋白乳酸饮料.本文介绍了绿豆酸奶的制作工艺及方法,通过对产品的分析证明绿豆酸奶是一种营养丰富乳酸菌饮料,含有人体必须的七种氨基酸,又含有医疗保健作用的药用成份黄酮、皂甙、鞣质,是一种较为理想的保健饮料.     

双歧酸奶定量干燥菌种的实验室研究

目的:对双歧酸奶定量干燥菌种的实验室研究进行初步探讨.方法:培养收获三种乳酸菌(双歧杆菌、乳杆菌、嗜热链球菌),并采用真空冷冻干燥技术进行处理,从而对干燥效果以及常温保存活性进行评价和分析.结果:三种乳酸菌的干燥菌粉活菌计数均大于1012个/g;凝乳试验显示次代菌种发酵时间明显低于第一代;混合发酵的凝乳中活菌计数均大于106个/ml,单一菌种发酵的凝乳中活菌计数超过1010个/ml;干燥菌种在37℃放置30 d后,活菌数仍可达108个/g以上.结论:双歧酸奶定量干燥菌种实验室研究结果令人满意,为开发双歧酸奶袋装式干燥菌种打下了基础.     

Scarce evidence of yogurt lactic acid bacteria in human feces after daily yogurt consumption by healthy volunteers

In a double-blind prospective study including 114 healthy young volunteers, the presence in human feces of the yogurt organisms Lactobacillus delbrueckii and Streptococcus thermophilus after repeated yogurt consumption (15 days) was analyzed by culture, specific PCR, and DNA hybridization of total fecal DNA. Detection of yogurt lactic acid bacteria in total fecal DNA by bacterial culture and PCR assay was consistently negative. DNA compatible with yogurt bacteria was found by hybridization experiments in only 10 (10.52%) of 96 individuals after consumption of fresh yogurt and in 2 (2.10%) of 96 individuals after consumption of pasteurized yogurt (P = 0.01).     

Dynamics of fecal microbiota in hospitalized elderly fed probiotic LKM512 yogurt

The comprehensive dynamics of intestinal microbiota including uncultured bacteria in response to probiotic consumption have not been well studied. The aims of this study were twofold: firstly to analyze the impact on intestinal microbiota of yogurt fermented by Bifidobacterium animalis subsp. lactis LKM512, Lactobacillus delbrueckii subsp. bulgaricus LKM1759, and Streptococcus thermophilus LKM1742 (LKM512 yogurt) and placebo fermented by these lactic acid bacterial strains without LKM512; and secondly to investigate the changes in intestinal microbiota that influence the concentration of PA, one of the beneficial metabolites produced by bacteria in the intestine. The LKM512 yogurt/placebo trial was performed in six hospitalized elderly patients (three men and three women with an average age of 80.3 years) and lasted seven weeks with the following schedule: pre-consumption for one week, LKM512 yogurt consumption for two weeks, washout period for two weeks, and placebo consumption for two weeks. The amount of ingested LKM512 yogurt or placebo was 100 g/day/individual. Fecal samples were analyzed using T-RFLP and real-time PCR. The T-RFLP patterns in five of the six volunteers were changed in a similar fashion by LKM512 yogurt consumption, although these patterns were individually changed following consumption of placebo. It was confirmed that B. animalis subsp. lactis was increased dramatically and Lactobacillus spp. tended to be decreased by LKM512 yogurt consumption. Some indigenous uncultured bacteria were increased and some decreased by LKM512 yogurt/placebo consumption. The similar changes in the intestinal microbiota of the elderly caused by consumption of the LKM512 yogurt were found to be influenced by the LKM512 strain itself, and not by the lactic acid bacteria in the yogurt. Moreover, this study suggests that the increase in intestinal PA concentrations caused by LKM512 yogurt consumption is probably dependent on the LKM512 strain colonizing the intestine.     

Scarce Evidence of Yogurt Lactic Acid Bacteria in Human Feces after Daily Yogurt Consumption by Healthy Volunteers

In a double-blind prospective study including 114 healthy young volunteers, the presence in human feces of the yogurt organisms Lactobacillus delbrueckii and Streptococcus thermophilus after repeated yogurt consumption (15 days) was analyzed by culture, specific PCR, and DNA hybridization of total fecal DNA. Detection of yogurt lactic acid bacteria in total fecal DNA by bacterial culture and PCR assay was consistently negative. DNA compatible with yogurt bacteria was found by hybridization experiments in only 10 (10.52%) of 96 individuals after consumption of fresh yogurt and in 2 (2.10%) of 96 individuals after consumption of pasteurized yogurt (P = 0.01).     

多菌种酸奶中活性乳酸菌的计数方法研究

利用4种选择性培养基MRS、MRS-山梨醇、MRS-5．2、Elliker,采用平板涂布法和倾注接种法。在蜡烛缸法（缺氧）、封口膜法（微氧）和供氧条件下对4种市售多菌种酸奶中保加利亚乳杆菌、嗜热链球菌、嗜酸乳杆菌、双歧杆菌进行选择性计数方法研究。结果表明：蜡烛缸法较能反映乳酸菌活菌数量的实际情况,封口膜法次之;在不同培养条件下4种选择性培养基对于乳酸菌活菌的计数都是灵敏的;而涂布法和倾注法接种活菌数有显著差异,倾注法要优于涂布法。     

Growth and activity of Bulgarian yogurt starter culture in iron-fortified milk

Bulgarian yogurts were manufactured and fortified with 8, 15 and 27 mg of iron kg(-1) of yogurt. The growth and acidifying activity of the starter culture bacteria Streptococcus thermophilus 13a and Lactobacillus delbrueckii subsp. bulgaricus 2-11 were monitored during milk fermentation and over 15 days of yogurt storage at 4 degrees C. Fortifying milk with iron did not affect significantly the growth of the starter culture during manufacture and storage of yogurt. Counts of yogurt bacteria at the end of fermentation of iron-fortified milks were between 2.1 x 10(10) and 4.6 x 10(10) CFU ml(-1), which were not significantly different from numbers in unfortified yogurts. In all batches of yogurt, the viable cell counts of S. thermophilus 13a were approximately three times higher than those of L. delbrueckii subsp. bulgaricus 2-11. Greater decrease in viable cell count over 15 days of storage was observed for S. thermophilus 13a compared to L. delbrueckii subsp. bulgaricus 2-11. Intensive accumulation of lactic acid was observed during incubation of milk and all batches reached pH 4.5 +/- 0.1 after 3.0 h. At the end of fermentation process, lactic acid concentrations in iron-fortified yogurts were between 6.9 +/- 0.4 and 7.3 +/- 0.5 g l(-1). The acidifying activity of starter culture bacteria in the control and iron-fortified milks was similar. There was no increase in oxidized, metallic and bitter off-flavors in iron-fortified yogurts compared to the control. Iron-fortified yogurts did not differ significantly in their sensorial, chemical and microbiological characteristics with unfortified yogurt, suggesting that yogurt is a suitable vehicle for iron fortification and that the ferrous lactate is an appropriate iron source for yogurt fortification.     

Reverse dot blot hybridization: A useful method for the direct identification of lactic acid bacteria in fermented food

Abstract A rapid method for a reliable and simultaneous identification of different lactic acid bacteria in fermented food has been developed. Various 16S and 23S rRNA-targeted, species-specific oligonucleotides were applied as capture probes in a non-radioactive reverse dot blot hybridization. A simple and fast DNA extraction method in combination with in vitro amplification of rRNA gene fragments enables the direct detection of typical starter organisms without any preceding enrichment or cultivation steps. Various lactic acid bacteria occurring in cheese, yogurt, sausages, sauerkraut and sourdough could be identified at the species level within 1 day.     

酸奶中乳酸菌镜检涂片的特殊染色法

报道了酸奶中乳酸菌涂片的特殊染色法-甲苯胺蓝染色法。涂片固定后,用0．2％甲苯胺蓝染色液染色2min,然后,用1％乙酸溶液脱色数秒,即可使涂片中的菌体着奥蓝色,而背景呈现白色或无色,菌体形状十分清晰。即使涂片很厚且不均,也可清晰看到菌体。便于菌体细胞的计数,形态观察及摄影．     

Dynamic metagenome-scale metabolic modeling of a yogurt bacterial community

Genome-scale metabolic models and flux balance analysis (FBA) have been extensively used for modeling and designing bacterial fermentation. However, FBA-based metabolic models that accurately simulate the dynamics of coculture are still rare, especially for lactic acid bacteria used in yogurt fermentation. To investigate metabolic interactions in yogurt starter culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, this study built a dynamic metagenome-scale metabolic model which integrated constrained proteome allocation. The accuracy of the model was evaluated by comparing predicted bacterial growth, consumption of lactose and production of lactic acid with reference experimental data. The model was then used to predict the impact of different initial bacterial inoculation ratios on acidification. The dynamic simulation demonstrated the mutual dependence of S. thermophilus and L. d. bulgaricus during the yogurt fermentation process. As the first dynamic metabolic model of the yogurt bacterial community, it provided a foundation for the computer-aided process design and control of the production of fermented dairy products.     

富锌产γ-氨基丁酸乳酸菌的筛选及初步鉴定

用含0.2%-0.25%Zn^2＋的MRS培养基从发酵酸菜、酸奶中分离出6株具富锌能力细菌,经形态和生理生化特性鉴定,初步判断菌株为乳酸菌。采用纸层析检测MRSG培养基培养的6株菌上清液后,初步确定有4株能产γ-氨基丁酸。以此4菌株基因组为模板PCR扩增到540 bp的片段,经测序和序列分析,证明是谷氨酸脱羧酶基因高度保守区域,初步证明谷氨酸脱羧酶基因的存在。此4株菌发酵液经HPLC检测和分析后,证实2号菌株能转化谷氨酸钠生成GABA,产量为4.8 g/L,在应用方面具有很大的潜力。     

Production of yogurt with enhanced levels of gamma-aminobutyric acid and valuable nutrients using lactic acid bacteria and germinated soybean extract

Yogurt with high levels of gamma-aminobutyric acid (GABA), free amino acids and isoflavones was developed using lactic acid bacteria (LAB) and germinated soybean extract. Fermented soya milk (GABA soya yogurt) produced with starter and substrate had the GABA concentration of 424.67 microg/gDW, whereas fermented milk produced by a conventional method had GABA less than 1.5 microg/gDW. The GABA soya yogurt also contained significantly high levels of free amino acids and isoflavones compared with other conventional yogurts. The results suggested that the Lactobacillus brevis OPY-1 and germinated soybean possessed a prospect to be applied in dairy and other health products with high nutritive values and functional properties.